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PubMed2004

Human immature germ cells and ejaculated spermatozoa contain aromatase and oestrogen receptors.

It is now well established that oestrogens play a part in germ cell function. These hormones are synthesised by the cytochrome P450 aromatase (P450 arom) and act via two kinds of receptor (ERalpha and ERbeta). Although the presence of aromatase and oestrogen receptors in mammalian testis is now well documented, the localisation of these proteins in human germ cells is not yet clear. The primary purpose of the current study was to look for the expression of aromatase and oestrogen receptors in human germ cells. Human immature germ cells were collected from semen samples with an excess of rounds cells (>20%) and purified spermatozoa were obtained after sedimentation on a discontinuous PureSperm gradient. Expression of aromatase and oestrogen receptors was determined by RT-PCR with specific primers, and by Western blot using monoclonal antibodies. RT-PCR products for aromatase, ERalpha and ERbeta were amplified from total RNA isolated from human germ cells and spermatozoa. We identified an ERalpha isoform variant that lacks exon 4 in human germ cells and visualised P450 arom as a single band of 49 kDa in germ cells, as we have already reported for human ejaculated spermatozoa. By Western blot, we identified two proteins for ERbeta at approximately 50 and approximately 60 kDa, which could correspond to the long and short forms of ERbeta formed from the use of alternative start sites. In human ejaculated spermatozoa, ERbeta protein was not detected, even though we could amplify mRNA. Using Western blot analysis and a monoclonal antibody specific for ERalpha, we detected two proteins in human immature germ cells: one of the expected size (66 kDa) and a second one of 46 kDa. In mature spermatozoa, only the 46 kDa band was observed and we speculate it may be related to the ERalpha isoform lacking exon I. In conclusion, we have identified P450 arom and ER proteins (full-length and variant) in human germ cells. Further studies are now required to elucidate the mechanism of action of oestrogens on human male germ cells, in terms of both genomic and 'non-genomic' pathways.

Adult↗

PubMed1985

Seminal biochemistry and sperm characteristics in infertile men with bacteria in ejaculate.

Sperm examination, quantitative sperm culture, citric acid, acid phosphatase, and fructose were assayed in three groups of men: fertile controls without significant bacteriospermia (group I), infertile men with significant bacteriospermia; idiopathic infertile men (group II), and infertile men with varicocele (group III). Level of significance of bacteriospermia was greater than or equal to 10(4) germs/ml of ejaculate. In group II, motility and typical morphology percentages were lower, independently of the degree and the nature of bacteriospermia. Incidence of pathogenic bacteria was higher than in group III and linked to the degree of bacteriospermia. Fructose was unaltered in the two groups of infected men. No modification of prostatic markers was observed in any groups, except in group II, where they decreased when bacteriospermia was lower than 10(5) germs/ml and when biological pattern of semen evoked chronic prostatitis. Thus, the presence of germs in ejaculate alters the motility and the typical morphology percentages but does not result in any obvious modifications of biochemical markers of prostate and seminal vesicles. For idiopathic infertile men, it is suggested that the quantitative criterion of pathogenic bacteriospermia is a germ count greater than or equal to 10(5)/ml.

Acid Phosphatase↗

PubMedDate not supplied

Synthesis of ether lipids and phosphatidylethanolamine by ejaculated human spermatozoa.

Ether lipids are the 1-O-alkyl derivatives of phospholipids. In contrast to nongerminal tissues where the plasma membrane content of ether lipids is low, over 40% of the phospholipids present in sperm plasma membranes are ether lipids. This study was undertaken to determine whether ejaculated human serm could synthesize ether lipids either through reacylation of 1-alkyl-sn-2-lysophosphatidylcholine or through direct incorporation of 1-hexadecanol into diacyl phosphatidylcholine or diacyl phosphatidylethanolamine. The ability of sperm to reacylate 1-acyl-sn-2-lysophosphatidylethanolamine was also assessed. In these experiments, freshly ejaculated sperm were unable to reacylate a phosphocholine lyso ether lipid with either palmitic (16:0) or docosahexaenoic (22:6) acids. In contrast, sperm readily incorporated both 16:0 and 22:6 into 1-acyl lysophosphatidylethanolamine. Similarly, sperm freely incorporated 1-hexadecanol into diacyl phosphatidylethanolamine thus forming a 1-alkyl phosphoethanolamine ether lipid. Diacyl phosphatidylcholine could not serve as a substrate in this reaction. It is apparent, based on these data, that human spermatozoa can directly synthesize phosphoethanolamine ether lipids that may subsequently undergo exhaustive methylation to form phosphocholine ether lipids.

Acylation↗

PubMedDate not supplied

Unaltered protein pattern/genital tract secretion marker levels in seminal plasma of highly viscous human ejaculates.

SDS-PAGE (12.5%) analysis and neutral alpha-glucosidase, fructose, and zinc level assessment were carried out in seminal plasma of 20 patients with highly viscous ejaculates and of 20 control subjects, with the aim to investigate the relations between high consistency of semen and epididymal, vesicular, and prostatic secretions. Very low sperm motility was observed in all the patients' ejaculates, both normo- and oligozoospermics. Protein patterns obtained in control and highly viscous semina showed similar protein bands, in the range of 10-100 kD. Furthermore, unaltered seminal neutral alpha-glucosidase, zinc, and fructose level were measured in the same specimens. These results indicated no impairment of epididymal, vesicular, and prostatic function in patients with hyperviscous semina, while their normal electrophoretic seminal protein profile suggested unaltered genital fluid interactions during the semen coagulation-liquefaction process.

Adolescent↗

PubMed2000

The method of assisted ejaculation affects the outcome of semen quality studies in men with spinal cord injury: A review.

The methods of penile vibratory stimulation (PVS) and rectal probe electroejaculation (EEJ) are widely used to obtain semen from anejaculatory men with spinal cord injury. This paper reviews evidence that the semen quality resulting from PVS versus EEJ is different, and these methods should be used as variables in studies of men with spinal cord injury. The following evidence is reviewed. PVS and EEJ stimulate ejaculation by different mechanisms. In humans and monkeys, semen quality is different when obtained by PVS versus EEJ of the same group of subjects. In neurologically intact men, EEJ results in low sperm motility. Markers of accessory gland function are different in ejaculates obtained by PVS versus EEJ. For scientific studies, it is recommended that whenever possible, antegrade specimens obtained by PVS be used. For clinical applications, such as obtaining sperm for assisted reproductive procedures, the choice is less critical, however, PVS offers advantages over EEJ.

Journal Article↗

PubMed2003

Evidence of aromatase localization in cytoplasmic droplet of human immature ejaculated spermatozoa.

Lytochrome P450 aromatase is a microsomal enzyme catalyzing the conversion of androgens to estrogens. P450arom expression has been demonstrated in testicular and epididymal sperm cells of several species but very limited data have been reported about maturating human germ cells. In this study, human spermatozoa with cytoplasmic droplet anomaly have been utilized to investigate aromatase immunolocalization in the immature germ cells of human ejaculate. Immunodetection has utilized a polyclonal antiserum as primary antibody, a biotinylated IgG as secondary antibody and then the avidin-biotin-peroxidase complex amplification followed by the diaminobenzidine staining. A strong immunoreaction was observed in the cytoplasmic droplets retained around the midpiece of immature spermatozoa and also in the descending droplets of late maturing sperm, while the other cellular components were unstained. Therefore, this investigation has demonstrated, for the first time, aromatase immunolocalization in residual cytoplasm of human ejaculated sperm, suggesting cytoplasmic droplets as possible estrogen biosynthesis sites during human sperm differentiation.

Aromatase↗

PubMed2006

[Functional expression of adenylyl cyclase and phosphodiesterase in ejaculated human spermatozoa].

OBJECTIVE: To compare the differences of expressions of adenylyl cyclase (AC) and phosphodiesterase (PDE) in ejaculated spermatozoa between healthy volunteers and the patients with asthenospermia. METHODS: Ejaculated spermatozoa were collected from healthy volunteers and the patients with asthenospermia. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect mRNA expression of AC and PDE subtypes in human spermatozoa. The concentrations of cAMP and cGMP in the samples were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared with healthy volunteers, expression of sAC mRNA and concentration of cAMP were significantly decreased in the patients with asthenospermia (P < 0.01) , while the expression of PDE4C mRNA was significantly increased at the same time (P <0.01). There were no marked differences in the expression of ACIII mRNA and concentration of cGMP between the two groups. CONCLUSION: The sAC down-regulation and PDE4C up-regulation are possible reasons for asthenospermia.

Adenylyl Cyclases↗

PubMed1991

[Analysis of the motility of bull sperm in fresh ejaculates using computer technology].

The activity of spermatozoa was measured in the fresh non-diluted ejaculates of 10 breeding bulls, using the HTM motility analyzer version 7. The average path speed was 83.6 microns.s-1, the average progressive speed was 48.2 microns.s-1 and the average straightness of the movement path was 58%. The spermatozoa were classified and it was found that most frequently they moved at a path speed of 60-80 microns.s-1 (28.2%) and at a progressive speed of 20-40 microns.s-1 (33.2%). The path straightness classes above 40% included evenly distributed numbers of spermatozoa; in classes with a less than 40% straightness the numbers of spermatozoa were much smaller. These data are characteristic of fresh undiluted bull ejaculates, suitable for artificial insemination.

Animals↗

PubMed1990

[The effect of the sperm count in the insemination dose and storage time on the fertilizing capability of the ejaculate of roosters].

Ejaculate of cooks was diluted by the Wishart and Davtjan diluents at a ratio of 1 : 1 and stored at a temperature of 4 degrees C during 2, 24, 48 and 72 hours before insemination of laying hens by the doses of 25, 50, 75, 100 and 150. 10(6) spermatozoa. With both diluents, egg fertility higher than 90 per cent was gained after 48 hours of keeping the ejaculate in sperm doses of 75 and 100 . 10(6) spermatozoa in the case of both diluents.

Animals↗

PubMedDate not supplied

Occurrence and potential importance of selected peptidases in bull ejaculates.

The zinc containing peptidases angiotensin-converting enzyme (ACE), neutral metalloendopeptidase (NEP), and leucine aminopeptidase (LAP) occur in bull ejaculates in high activities. These enzyme activities are in close correlation with some routineously determined semen parameters. These ejaculat parameters are used for quality classification and selection of ejaculates and are according to long term experience in good correlation to the male fertility. Semen quality related correlations could not be found for the dipeptidyl aminopeptidase (DPIV) and the aminopeptidase A (APA). Origin and possible function of the zinc containing peptidases in the semen are examined and discussed.

Animals↗

PubMed1988

[Bacteriology of the ejaculate--a useful study?].

In a retrospective study we analyzed the results of 318 bacteriologic tests of ejaculates from 175 patients with suspected infections of the prostate, epididymis and urethra. Only 55% of all bacteriologic tests were positive, and the bacteria most frequently found were those normally present in the anterior male urethra, enterococci and coagulase-negative staphylococci. These results indicate that bacteriologic testing of the ejaculate does not yield such reliable results as the segmented urine culture technique and the examination of expressed prostatic secretions.

Adolescent↗

PubMed1975

Absorption activity of the blood group substances in various fractions of split ejaculates from man.

Antigens of the ABO system have been determined in the various fractions of split ejaculates from three A and two O (H) men who were secretors. The activity of the antigen was determined by a hemagglutination inhibition test. Within a split ejaculate the various fractions contained almost the same antigen activity, indicating that the antigens were secreted from all the glands contributing to the seminal plasma.

ABO Blood-Group System↗

PubMed1987

Passage of exogenous L-thyroxine and triiodothyronine into bovine ejaculate.

The metabolic effects of thyroxine (T4) and triiodothyronine (T3) on spermatozoa metabolism and male anatomy have been demonstrated. The metabolic effects of T3 and T4 could affect the physiologic characteristics of the spermatozoa. There are little data on the passage of T4 and T3 into the ejaculate from blood. The passage of exogenous T4 and T3 from the blood into semen was measured after T4 (45 mg) or T3 (37.5 mg) was injected IV into 8 bulls. Blood and electroejaculate were obtained simultaneously at 20, 40, 60, 120, and 180 minutes and 24 hours after bulls were injected to determine T3 and T4 concentrations compared with base-line values. Blood T3 and T4 concentrations were increased (P less than 0.05) at 20 minutes after bulls were injected (1.1 +/- 0.25 to 598 +/- 76.3 ng/ml and from 66 +/- 5 to 1,318 +/- 105 ng/ml, respectively). Seminal concentrations of T4 were unchanged until 120 minutes after bulls were injected, when they increased (P less than 0.05) from less than 1.2 ng/ml to 4.7 +/- 1.9 ng/ml. However, seminal concentrations of T3 were increased (P less than 0.05) from less than 0.1 ng/ml to 0.5 +/- 0.2 ng/ml at 20 minutes and to 12.5 +/- 2.9 ng/ml at 120 minutes after bulls were injected. It was concluded that exogenous thyroid hormones passed into the ejaculate from blood, with T3 passing faster than T4.

Animals↗

PubMed1975

[Mycoplasma prostatourethritis. The improtance of immunoelectrophoresis of the ejaculate for the diagnosis].

The secretions expessed from the prostate and/or urethral smears from 846 patients of a prostatitis clinic were examined microbiologically. With negative findings in 62.9%, which suggested a high proportion of patients with a vegetative urogenital syndrome, mycoplasma could be isolated in 9.9% of the entire collective. In the cases with mycoplasma in the expressed prostatic secretion, a close, statistically significant correlation to the degree of clinical complaint and the mumber of leukocytes in the secretion was established. Immunoelectrophoresis of the ejaculate was carried out in 20 subjects in the mycoplasma group. Although unequivocal statement are not yet possible because of the small number of cases, there is a statistically striking trend towards infection with urea-splitting mycoplasma when the immunoelectrophoresis of the ejaculate is unchanged, which might indicate the induction of a predominantly localised urethritis due to this strain.

Humans↗

PubMed1996

Chlamydia trachomatis antibodies in serum and ejaculate of male patients without acute urethritis.

In a prospective study, the prevalence of specific antibodies against Chlamydia trachomatis in serum and seminal plasma evaluated by a genus specific immunofluorescence test in 101 men without acute urethritis was investigated. The results were compared to the clinical diagnosis, cell culture of urethral swabs, demonstration of DNA particles by Polymerase Chain Reaction (PCR) in the ejaculate and signs of genital inflammation by counting peroxidase-positive leukocytes and elastase level in semen. The objective was to evaluate the significance of chlamydial antibodies in genital infection. Serum specific IgG and IgA antibodies were found in 26% and 15%, respectively; seminal IgG and IgA antibodies were present in 6% and 7%, respectively. Serum specific antibodies were not associated with the clinical diagnosis of infection nor with C. trachomatis cell culture, PCR findings, peroxidase positive leukocytes or PMN-elastase level. It is concluded that serum antibodies are not useful in detecting a chlamydial infection. Seminal plasma antibodies were not correlated with the clinical diagnosis of infection, positive cell culture, PMN-elastase levels and leukocytes in semen. However, a significant correlation was found for positive PCR in the ejaculate (p < 0.001 for IgG, p < 0.05 for IgA, p < 0.001 when combined). Though seminal antibodies may be more useful in detecting ascended or occult chlamydial infection, their significance remains unclear, their absence does not exclude chlamydial infection. In particular, the biological significance of locally derived IgA needs further evaluation.

Acute Disease↗

PubMedDate not supplied

[Iron and nonheme iron protein metabolism in ejaculates with varying degrees of fertility].

Spermoplasm of 21 fertile and 96 subfertile males was examined for Fe, transferrin and scaferrin the concentrations of which were compared to 14 morphofunctional and physicochemical parameters of ejaculate fertility. Fe and non-hemic spermoplasm ferroproteins are involved in male reproduction: participate in ejaculate thinning, in formation of viscosity and sperm pH, are important for normal spermatogenesis and intercellular interactions. The findings can be applied in diagnosis of male subfertility and control of ongoing treatment, hold promise for improvement of fertilizing ability of the sperm used for artificial insemination and extracorporeal fertilization.

Adult↗

PubMed2026

Combined effects of urine exposure and cryopreservation on sperm quality: an in vitro study of retrograde ejaculation.

Sperm quality influences fertility and offspring health through both genomic inheritance and epigenetic inheritance. Thus, for use in clinical-assisted reproductive technology (ART), spermatozoa must have optimal genomic and epigenetic structures. In patients with retrograde ejaculation, spermatozoa are usually recovered from urine and then cryopreserved for ART. However, the effects of urine exposure and subsequent freeze-thaw cycles on sperm quality remain unclear. This is particularly true for epigenetic changes and their underlying mechanisms. In this study, we examined how different durations of urine exposure (10 min and 40 min) followed by freeze-thaw cycles affected sperm motility, DNA integrity, and methylation levels of imprinting genes (H19-imprinted maternally expressed transcript [ H19 ], mesoderm-specific transcript [ MEST ], and the transposable element Alu [ Alu ]). As the duration of urine exposure increased, sperm motility (median [interquartile range]) decreased from 48.0% (39.0%-52.5%) to 1.0% (1.0%-5.0%), the DNA fragmentation index (DFI; median [interquartile range]) increased from 12.0% (9.3%-19.9%) to 23.5% (13.9%-33.9%), the MEST methylation level (mean ± standard deviation [s.d.]) increased from 3.8% ± 1.5% to 11.5 ± 1.2%, and the H19 methylation level (mean ± s.d.) decreased from 86.9% ± 0.9% to 82.1% ± 0.5%. The freeze-thaw process further reduced sperm motility, while the DFI and methylation levels of MEST and H19 did not significantly change. The Alu methylation level remained stable. These findings demonstrate that urine exposure affects sperm motility, DNA integrity, and methylation levels of some imprinting genes. These effects intensify over time. In contrast, the freeze-thaw process impacts only sperm motility. In clinical practice, minimizing exposure to urine might improve sperm quality.

Humans↗

PubMed1989

Effects of psychotropic drugs on human erection and ejaculation.

Evidence concerning pharmacological effects on human sexuality suggests that dopaminergic receptor activation may be associated with penile erection. Erection also appears to involve inhibition of alpha-adrenergic influences and beta-adrenergic stimulation plus the release of a noncholinergic vasodilator substance, possibly vasoactive intestinal peptide. Ejaculation appears to be mediated primarily by alpha-adrenergic fibers. Serotonergic neurotransmission may inhibit the ejaculatory reflex. An understanding of the neurobiological substrate of human sexuality may assist clinicians in choosing psychotropic agents with minimal adverse effects on sexual behavior and may also contribute to the development of pharmacological interventions for sexual difficulties.

Ejaculation↗